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Transfection - DNA Transfection
DNA Transfection
The addition of foreign DNA put into a cultured mammalian cell is the process of transfection. It is the basis of gene therapy and modern genetic research. Experiments are usually performed using cloned DNA containing coding sequences and control regions (promoters, etc) which will test whether the DNA will be expressed. Since the cloned DNA may have been extensively modified, transfection is often used to test whether a particular modification affects the function of a gene.
Cell transfection, can be accomplished chemically, biologically, or mechanically. Current methods include eletroporation, use of a virus vector, lipofection, gene guns, and microinjection.
Electroporation uses electricity to increase the permeability of the cell membrane, allowing the DNA to pass easily inside. This method is good for infecting large numbers of cells at once.
A virus vector causes little cell damage and can deliver a wide variety of DNA.
Lipofection, which delivers DNA by fusing to the cell wall and allowing its DNA payload to be absorbed into the cell.
Microinjection and gene guns carry a risk of destroying the cell, but they are both excellent ways to deliver DNA to a specified target.
Transfection is either transient or stable in nature. It is transient when foreign DNA is introduced, but is eliminated by the cell prior to or during mitosis. Stable transfection is rare but occurs when foreign DNA is introduced and enters the cell nucleus, adding to or replacing a portion of the cell's native DNA. This transforms the cell so that the DNA change is duplicated during mitosis.
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